Compositions comprising polyunsaturated fatty acid monoglycerides, derivatives thereof and uses thereof

ABSTRACT

There are provided various compounds and compositions comprising polyunsaturated fatty acid monoglycerides and derivatives thereof. These compounds and compositions can be useful as cancer chemopreventive agents. They can also be useful for enhancing solubility of various active agents and enhancing their bioavailability.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is divisional application of U.S. Ser. No.12/536,519 filed on Aug. 6, 2009, that is continuation-in-part of PCTinternational patent application No. PCT/CA2008/000530 filed on Mar. 19,2008, which claims priority on U.S. provisional application No.60/895,795 filed on Mar. 20, 2007. These applications are incorporatedherein by reference in their entirety.

FIELD OF THE DISCLOSURE

The present disclosure relates to the field of medicinal chemistry. Moreparticularly, it relates to polyunsaturated fatty acid monoglyceridecompounds and derivatives thereof. It also relates to compositions whichcomprise such compounds. There is also provided a method for enhancingthe bioavailability of an active agent and a method for enhancingsolubility of such an active agent.

BACKGROUND OF THE DISCLOSURE

The primary targets for any dosage formulation is to deliver thenecessary concentration of an active drug to the site of action toelicit the desired therapeutic response and to maintain an effectiveconcentration of the drug for a sufficient period to achieve efficacioustreatment. Oral administration is generally preferred but is frequentlydependent upon the bioavailability of the active form of the drug, i.e.,the rate and extent that the active form of the drug appears from thedosage form in the systemic circulation. Bioavailability is affected bythe drug's physical chemical properties, such as pKa, water solubility,oil solubility and stability, as well as its absorption, distribution,metabolism and excretion. It is well known that water insoluble drugsare not generally available for absorption through intestinal lumin andoil insoluble drugs are generally unable to pass across intestine cellmembranes into systemic circulation (S. H. Yalkowsky, “DRUGS AND THEPHARMACEUTICAL SCIENCES: TECHNIQUES OF SOLUBILIZATION OF DRUGS,” MarcelDekker, Inc., Vol. 12, 1981). Proper formulations can improve thebioavailability of a drug.

SUMMARY OF THE DISCLOSURE

According to one aspect there are provided compounds of formulas (I),(II), (III), and (IV):

wherein

X₁ is O, NH, or S;

X₂ is O, NH, or S;

X₃ is O, NH, or S;

R₁ and R₂ each independently represents —H, —C(O)NH₂, —S(O)NH₂,—S(O)₂NH₂, —C1-C22 (oxy)alkyl, —C1-C22 alkyl, —C1-C22 (hydroxy)alkyl,—C1-C22 (amino)alkyl, —C1-C22 (halo)alkyl, —C3-C22 alkenyl, —C3-C22alkynyl, —(C3-C7) cycloalkyl unsubstituted or substituted with at leastone substituent chosen from C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22alkynyl, —C6-C12 aryl, —C7-C22 (aryl)alkyl, —C8-C22 (aryl)alkenyl,—C8-C22 (aryl)alkynyl, three- to seven-membered non-aromatic heterocycleunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, five- toseven-membered aromatic heterocycle unsubstituted or substituted with atleast one substituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and—C2-C22 alkynyl, —(CH₂)_(n)amino acid wherein the amino acid isconnected through its alpha carbon atom, —(CH₂)_(n)peptide wherein thepeptide is connected through the alpha carbon atom of one of its aminoacids, —CH₂OR₅, —C(O)R₅, —C(O)OR₅, —C(O)NR₅, —P(O)(OR₅)₂, —S(O)₂NHR₅,—SOR₅, —S(O)₂R₅, -arylP(O)(OR₅)₂, a sugar, or a sugar phosphate

or R₁ and R₂ are joined together so as to form a five- to seven-memberednon-aromatic heterocycle unsubstituted or substituted with at least onesubstituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22alkynyl, a phosphate, sulfate carbonyl group, or a thiocarbonyl imine;

R₅ is —H, —C1-C22 alkyl, —(C3-C7) cycloalkyl, —C1-C22 (halo)alkyl,—C6-C12 aryl, —C2-C22 alkenyl, —C2-C22 alkynyl, —C7-C22 (aryl)alkyl,—C8-C22 (aryl)alkenyl, —C8-C22 (aryl)alkynyl, —C1-C22 (hydroxy)alkyl,—C1-C22 alkoxy, —C1-C22 (amino)alkyl, a —(C3-C7) cycloalkylunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, a three- toseven-membered non-aromatic heterocycle unsubstituted or substituted atleast one substituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and—C2-C22 alkynyl, a three- to seven-membered aromatic heterocycleunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, a —(CH₂)_(n)aminoacid wherein the amino acid is connected to the compound through itsalpha carbon atom, a —(CH₂)_(n)peptide wherein the peptide is connectedto the compound through the alpha carbon atom of one of its amino acids,a sugar or a sugar phosphate; and

n is an integer having a value of 0, 1, 2, 3, or 4,

and pharmaceutically acceptable salts thereof.

According to another aspect there are provided compounds of formulas(V), (VI), (VII), (VIII), (IX), (X), (XI), (XII), (XIII), (XIV) or (XV):

X₁ is O, NH, or S;

X₂ is O, NH, or S;

X₃ is O, NH, or S;

R₃ and R₄ each independently represents —H, —C(O)NH₂, —S(O)NH₂,—S(O)₂NH₂, —C1-C22 (oxy)alkyl, —C1-C22 alkyl, —C1-C22 (hydroxy)alkyl,—C1-C22 (amino)alkyl, —C1-C22 (halo)alkyl, —C3-C22 alkenyl, —C3-C22alkynyl, —(C3-C7) cycloalkyl unsubstituted or substituted with at leastone substituent chosen from C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22alkynyl, —C6-C12 aryl, —C7-C22 (aryl)alkyl, —C8-C22 (aryl)alkenyl,—C8-C22 (aryl)alkynyl, three- to seven-membered non-aromatic heterocycleunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, five- toseven-membered aromatic heterocycle unsubstituted or substituted with atleast one substituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and—C2-C22 alkynyl, —(CH₂)_(n)amino acid wherein the amino acid isconnected through its alpha carbon atom, —(CH₂)_(n)peptide wherein thepeptide is connected through the alpha carbon atom of one of its aminoacids, —CH₂OR₅, —C(O)R₄, —C(O)OR₄, —C(O)NR₄, —P(O)(OR₅)₂, —S(O)₂NHR₅,—SOR₅, —S(O)₂R₅, -arylP(O)(OR₅)₂, a sugar, or a sugar phosphate,

or R₃ and R₄ are joined together so as to form a five- to seven-memberednon-aromatic heterocycle unsubstituted or substituted with at least onesubstituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22alkynyl, a phosphate, sulfate carbonyl group, or a thiocarbonyl imine;

R₅ is —H, —C1-C22 alkyl, —(C3-C7) cycloalkyl, —C1-C22 (halo)alkyl,—C6-C12 aryl, —C2-C22 alkenyl, —C2-C22 alkynyl, —C7-C22 (aryl)alkyl,—C8-C22 (aryl)alkenyl, —C8-C22 (aryl)alkynyl, —C1-C22 (hydroxy)alkyl,—C1-C22 alkoxy, —C1-C22 (amino)alkyl, a —(C3-C7) cycloalkylunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, a three- toseven-membered non-aromatic heterocycle unsubstituted or substituted atleast one substituent chosen from —C1-C22 alkyl, —C2-C22 alkenyl, and—C2-C22 alkynyl, a three- to seven-membered aromatic heterocycleunsubstituted or substituted with at least one substituent chosen from—C1-C22 alkyl, —C2-C22 alkenyl, and —C2-C22 alkynyl, a —(CH₂)_(n)aminoacid wherein the amino acid is connected to the compound through itsalpha carbon atom, a —(CH₂)_(n)peptide wherein the peptide is connectedto the compound through the alpha carbon atom of one of its amino acids,a sugar or a sugar phosphate; and

n is an integer having a value of 0, 1, 2, 3, or 4;

and pharmaceutically acceptable salts thereof.

The previously mentioned compounds can be used separately or in amixture (or composition) of at least two of them (for example 2, 3, 4, 5or 6 of them). Thus, the present disclosure also includes compositionscomprising at elast two compounds of the present disclosure,

According to another aspect, there is provided a composition comprisingat least one compound chosen from compounds of formulas I, II, III, IV,V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV; and at least oneactive agent.

According to another aspect, there is provided a method for solubilizinga lipophilic active agent. The method comprises mixing the lipophilicactive agent with at least one compound chosen from compounds offormulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV,and XV. The method can further comprise mixing a lipid with the at leastone compound and the lipophilic active agent.

According to another aspect, there is provided a method for enhancingthe solubility of at least one active agent in a lipid or a lipidformulation. The method comprises mixing the at least one active agent,and at least one compound chosen from compounds of formulas I, II, Ill,IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV. The method cancomprise mixing together the at least one active agent and the at leastone compound so as to obtain a composition and then, mixing the lipidand the composition or the method can comprise mixing together the lipidand the at least one compound so as to obtain a composition and then,mixing the at least one active agent and the composition. For example,the lipid can be chosen from compounds of formulas I, II, III, IV, V,VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV or the lipidformulation can comprise at least one compound chosen from compounds offormulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV,and XV.

According to another aspect, there is provided a method for enhancingbioavailability of at least one active agent. The method comprisesmixing the at least one active agent with at least one compound chosenfrom compounds of formulas I, II, Ill, IV, V, VI, VII, VIII, IX, X, XI,XII, XIII, XIV, and XV.

For example, the compounds and compositions of the present disclosurecan be used for treating cancer (for example breast cancer, lung cancer,prostate cancer, colon cancer).

For example, the compounds and compositions of the present disclosurecan be used for enhancing bioavailability of at least one active agent.

According to another aspect, there is provided a method for inhibitingtumor growth, inhibiting tumor cell proliferation, or reducing tumorgrowth, in vitro or in vivo, The method comprises contacting the tumorwith an effective amount of at least one compound or composition aspreviously described.

According to another aspect, there is provided a method forchemopreventing cancer comprising administering to a subject aneffective amount of at least one compound or composition as previouslydefined. Such a cancer can be lung cancer, prostate cancer, breastcancer, or colon cancer.

According to another aspect, there is provided a method for reducingtumor growth in a subject comprising administering to the subject aneffective amount of at least one compound or composition as previouslydefined.

According to another aspect there is provided a method for treatingcancer (for example breast cancer, lung cancer, prostate cancer, coloncancer) comprising administering to the subject in need thereof aneffective amount of at least one compound chosen from compounds offormulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV,and XV.

According to another aspect there is provided a method for treatingcancer (for example breast cancer, lung cancer, prostate cancer, coloncancer) comprising administering to the subject in need thereof aneffective amount of at least one active agent and an effective amount ofat least one compound of the present disclosure. For example, acomposition comprising an effective amount of the at least one activeagent and an effective amount of at least one compound of the presentdisclosure can be administered. Alternatively, the effective amount ofthe at least one active agent and the effective amount of the at leastone compound can be administered separately. For example, the activeagent can be curcumin.

According to another aspect there is provided a method for inhibitingtumor growth, inhibiting tumor cell proliferation, or reducing tumorgrowth, in vitro or in vivo, comprising administering an effectiveamount of at least one active agent and an effective amount of at leastone compound of the present disclosure. For example, a compositioncomprising an effective amount of the at least one active agent and aneffective amount of at least one compound of the present disclosure canbe administered. Alternatively, the effective amount of the at least oneactive agent and the effective amount of the at least one compound canbe administered separately.

According to another aspect, there is provided a method for enhancingbioavailability of at least one active agent. The method comprisesmixing the at least one active agent with at least one compound of thepresent disclosure.

According to another aspect, there is provided a method for enhancingbioavailability of at least one active agent. The method comprisesadminitering to a subject an effective amount of the at least one activeagent and an effective amount of at least one compound of the presentdisclosure. For example, a composition comprising an effective amount ofthe at least one active agent and an effective amount of at least onecompound of the present disclosure can be administered. Alternatively,the effective amount of the at least one active agent and the effectiveamount of the at least one compound can be administered separately.

According to another aspect, there is provided a method for enhancingbioavailability of at least one lipophilic active agent, natural productor natural crude extract. The method comprises adminitering to a subjectan effective amount of the at least one lipophilic active agent, naturalproduct or natural crude extract and an effective amount of at least onecompound of the present disclosure. For example, a compositioncomprising an effective amount of the at least one lipophilic activeagent, natural product or natural crude extract and an effective amountof the at least one compound of the present disclosure can beadministered. Alternatively, the effective amount of the at least onelipophilic active agent, natural product or natural crude extract andthe effective amount of the at least one compound can be administeredseparately.

According to another aspect, there is provided a method for enhancingbioavailability of at least one active agent present in at least oneoil. The method comprises adminitering to a subject an effective amountof the at least one oil and an effective amount of at least one compoundof the present disclosure. For example, a composition comprising aneffective amount of the at least one oil and an effective amount of atleast one compound of the present disclosure can be administered.Alternatively, the effective amount of the at least one oil and theeffective amount of the at least one compound can be administeredseparately.

It was found that the compounds and compositions previously mentionedcan be useful as bioavailability enhancers of various types of activeagents.

It was also found that the compounds and compositions previouslymentioned can enhance the solubility of an active agent such as alipophillic active agent. It was observed that active agents can be moreeasily solubilized in the compounds and compositions previously definedthan in the usual lipids or lipid formulations. Thus, the active agentsthat need to be dissolved in a lipid or a lipid formulation can bedissolved in the compounds or compositions as those previously definedinstead than in other conventional lipids or lipid formulations.Alternatively, such active agents can be mixed with at least onecompound or composition as previously defined in order to obtain anothercomposition and then the other composition can be dissolved in a lipidor a lipid formulation. In such a case, the overall solubility of theactive agent in the lipid or lipid formulation is enhanced.

According to another aspect, there is provided a method for treating aninflammatory disease comprising administering to a subject in needthereof an effective amount of at least one active agent and aneffective amount of at least one compound of the present disclosure. Forexample, a composition comprising an effective amount of the at leastone active agent and an effective amount of at least one compound of thepresent disclosure can be administered. Alternatively, the effectiveamount of the at least one active agent and the effective amount of theat least one compound can be administered separately.

BRIEF DESCRIPTION OF THE FIGURES

Further features and advantages will become more readily apparent fromthe following description of specific embodiments as illustrated by wayof examples in the appended figures wherein:

FIG. 1 represents an in vitro assay of a composition according to anexample, wherein the assay was carried out on A549 human cancer cellline;

FIG. 2 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on A549 human cancercell line;

FIG. 3 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on A549 human cancercell line;

FIG. 4 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on PC3 human cancercell line;

FIG. 5 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on PC3 human cancercell line;

FIG. 6 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on PC3 human cancercell line;

FIG. 7 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on HCT-15 humancancer cell line;

FIG. 8 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on HCT-15 humancancer cell line;

FIG. 9 represents an in vitro assay of a composition according toanother example, wherein the assay was carried out on HCT-15 humancancer cell line;

FIG. 10 represents a comparative solubility study ofbis-demethoxycurcumin, demetoxycurcumin, curcumin and total curcuminoidsin two different solvents which are a fish oil and a compositionaccording to another example;

FIG. 11 represents a comparative in vivo absorption study of twodifferent formulations containing docosahexaenoic acid (DHA) which are afish oil and a composition according to another example;

FIG. 12 represents a comparative human absorption cross-over study oftwo different compositions containing docosahexaenoic acid (DHA) whichare a fish oil and a composition according to another example;

FIG. 13 represents a comparative human absorption cross-over study oftwo different compositions containing omega-3 docosapentaenoic acid(DPAω3) which are a fish oil and a composition according to anotherexample; and

FIG. 14 represents an in vitro assay of a composition according to anexample, wherein the assay was carried out on human THP-1 monocyte cell.

DETAILED DESCRIPTION OF THE DISCLOSURE

Further features and advantages of the previously-mentioned compoundswill become more readily apparent from the following description ofnon-limiting examples.

The compounds and compositions of the present disclosure can also beeffective so as to enhance solubility of various active agents. They canalso be used as a emulsifier alone, emulsifier in a self-emulsifyingdrug delivery systems (SEDDS), self-microemulsifying drug deliverysystems (SMEDDS) and selfemulsifying oil formulations (SEOF) forimproved oral delivery of a lipophilic active agent, natural product ornatural crude extract. They can also be used in combination with a1-monoglyceride, 2-monoglyceride, 1,2-diglyceride, 1,3-diglyceride,triglyceride, free fatty acid, phospholipid and pharmaceuticallyacceptable salts thereof. For example, they can also be used incombination with vegetable oil, fish oil, seal oil, microalgae oil,krill oil and crustacean oil (ex. shrimp oil). According to anotherexample, they can also be used in combination with oil hydrolysate fromvegetable oil, fish oil, seal oil, microalgae oil, krill oil andcrustacean oil (ex. shrimp oil). According to another example they canbe used in combination with proteins hydrolysate from vegetable, animaland marine source. Moreover, they can also be used to form micelle orliposome for a drug delivery system.

According to another example, the active agent can be a lipid or anhydrolysate thereof, or a protein or an hydrolysate thereof.

According to another example, the active agent can be chosen from afatty acid, a salt thereof, an ester thereof (for example amonoglyceride, a diglyceride, or a triglyceride), and mixtures thereof.

According to another example, the active agent can be a fatty acid or aderivative thereof (for example an C1-C6 ester (C1-C6 being the amountof carbon atoms in the “alcohol” portion of the ester) of a fatty acidsuch as an ethyl ester) or a pharmaceutically acceptable salt thereof.

According to another example, the active agent can be a polyunsaturatedfatty acid chosen from arachidonic acid, ω3-arachidonic acid,alpha-linolenic acid, conjugated linoleic acid, linoleic acid,gamma-linolenic acid, dihomo-gamma-linolenic acid, stearidonic acid,eicosapentaenoic acid, ω3-docosapentaenoic acid, ω6-docosapentaenoicacid, docosahexaenoic acid, monoglycerides thereof, diglyceridesthereof, triglycerides thereof, phospholipids thereof, and saltsthereof.

According to another example, the active agent can be a compound chosenfrom compounds previously defined.

According to another example, the active agent can be chosen from any ofthe following therapeutic class: analgesic, anesthetic,anti-Alzheimer's, anti-asthma agent, anti-Parkinsonism, antiallergic,antianginal, antiarrhythmic, antiarthritic, antiasthmatic,antibacterial, antibiotic, anticancer, anticoagulant, antidepressant,antidiabetic, antiemetic, antiepileptic, antifungal, antiglaucoma,anti-gout, antihistamine, antihyperprolactinemia, antihypertensive,antiinflammatory, antimigraine, anti-neoplastic, antiobesity,antiparasitic, anti-protozoal, anti-phyretics, antipsoriatic,antipsychotic, antithrombotic, antiulcer, antiviral, anxiolytic, benignprostatic hypertrophy, bronchodilator, calcium metabolism, cardiotonic,cardiovascular agent, chelator & antidote, chemopreventive agent,contraception, diuretic, dopaminergic agent, gastrointestinal agent,gastroprokinetic, hematopoiesis, hemophilia, hormone, hormonereplacement therapy, hypnotic, hypocholesterolemic, hypolipidemic,immunomodulator, immunostimulant, immunosuppressant, lipid regulatingagent, male sexual dysfunction, multiple sclerosis, muscle relaxant,neuroleptic, nootropic, osteoporosis, phytoestrogen, plateletaggregation inhibitor, prostaglandin, radioenhencer for radiotherapy,relaxant and stimulant, respiratory distress syndrome, urinaryincontinence, vasodilator, vitamin/nutritional, vulnerary and xanthine.Active agents belonging to these classes can be used in the previouslymentioned compositions.

According to one example, the active agent can be a chemopreventiveagent. The chemopreventive agent can be, for example, chosen from13-cis-retinoic acid, 9-cis retinoic acid, anetholtrithione, arzoxifenehydrochloride, aspirin, bexarotene, biaxin (clarithromycin), budesonid,calcium, celecoxib, curcumin, DFMO, DHEA (Dehydroepiandrosterone),fenretinide, indole-3-carbinol, l-perillyl alcohol, lycopene, oltipraz,phenethyl isothiocyanate (PEITC), piroxicam, raloxifen, selenium, soyisoflavones, sulindac, tamoxifen, 4-hydroxy-tamoxifen, citrate, teapolyphenols, ursodiol, vitamin D and analogs, and zileuton.

According to another example, the active agent can be an antibacterialagent. Non-limitative examples of antibacterial agents are RV-11,carumonam, daptomycin, fosfomycin trometamol, isepamicin, micronomicinsulfate, miokamycin, mupirocin, netilimicin sulfate, teicoplanin,apalcillin sodium, arbekacin, aspoxicillin, astromycin sulfate,azithromycin, aztreonam, biapenem, cefbuperazone sodium, cefcapenepivoxil, cefdinir, cefditoren pivoxil, cefepime, cefetamet pivoxil HCl,cefixime, cefmenoxime HCl, cefminox, sodium, cefodizime sodium,cefonicid sodium, cefoperazone sodium, ceforanide, cefoselis, cefotetandisodium, cefotiam HCl, cefozopran HCl, cefpimizole, cefpiramide sodium,cefpirome sulfate, cefpodoxime proxetil, cefprozil, cefsoludin sodium,ceftazidime, cefteram pivoxil, ceftibuten, ceftizoxime sodium,ceftriaxone sodium, cefuroxime axetil, cefuzonam sodium, clarithromycin,dalfopristin, dirithromycin, doripenem, ertapenem sodium, erythromycinacistrate, flomoxef sodium, flurithromycin ethylsuccinate, fropenam,imipenem/cilastatin, lenampicillin HCl, loracarbef, meropenem,moxalactam disodium, panipenem/betamipron, quinupristin, rifabutin,rifamixin, rifapentine, rifaximin, rokitamycin, roxithromycin,sultamycillin tosylate, tazobactam sodium, telithromycin, temocillindisodium, tigecycline, balafloxacin, ciprofloxacin, enoxacin,fleroxacin, gatilfloxacin, gemifloxacin mesilate, grepafloxacin,levofloxacin, linezolid, lomefloxacin, moxifloxacin HCl, nadifloxacin,norfloxacin, ofloxacin, pazufloxacin, pefloxacin mesylate,prulifloxacin, rufloxacin hydrochloride, sparfloxacin, taurolidine,temafloxacin hydrochloride, tosufloxacin, trovafloxacin mesylate,rodimoprin, ACWY meningoccal PS vaccine, MCV-4, h influenzae b vaccine,h influenzae b vaccine, meningitis b vaccine, meningococcal vaccine,oral cholera vaccine, pneumococcal vaccine, and vi polysaccharidetyphoid vaccine

According to another example, the active agent can be an antifungalagent. The antifungal agent can be, for example, chosen from interferongamma-n1, anidulafungin, caspofungin acetate, micafungin sodium,amorolfine hydrochloride, butoconazole, ciclopirox, olamine, cloconazoleHCl, eberconazole, fenticonazole nitrate, fluconazole, flutrimazole,fosfluconazole, itraconazole, ketoconazole, lanoconazole, luliconazole,naftifine HCl, neticonazole HCl, oxiconazole nitrate, posaconazole,sertaconazole nitrate, sulconazole nitrate, terconazole Gyno,tioconazole, voriconazole, butenafine hydrochloride, liranaftate, andterbinafine hydrochloride

According to another example, the active agent can be an antiviralagent. Non-limitative examples of antiviral agents are: immunoglobulinintravenous, interferon alfa, interferon alfa-2b, interferon alfa-n3,interferon alfacon-1, interferon beta, palivizumab, peginterferonalfa-2a, peginterferon alfa-2b, resp syncytial virus IG, thymalfasin,interferon alfa-n1, enfuvirtide, zanamivir, delavirdine mesylate,efavirenz, foscarnet sodium, imiquimod, nevirapine, propagermanium,rimantadine HCl, oseltamivir, abacavir sulfate, acyclovir, adefovirdipivoxil, cidofovir, didanosine, emtricitabine, entecavir, epervudine,famciclovir, ganciclovir, inosine pranobex, lamivudine, penciclovir,sorivudine, stavudine, tenofovir disoproxil fumarate, valaciclovir HCl,valganciclovir, zalcitabine, zidovudine, amprenavir, atazanavir,darunavir, fomivirsen sodium, fosamprenevir, indinavir sulfate,lopinavir, neflinavir mesylate, ritonavir, saquinavir mesylate,tipranavir, MR vaccine, anti-Hep B immunoglobulin, attenuated chickenpox vaccine, hepatitis A and B vaccine, hepatitis B vaccine, hepatitis avaccine, inact hepatitis a, influenza vaccine, influenza virus (live),rotavirus vaccine, rubella vaccine, varicella virus vaccine, and zostervaccine live.

According to another example, the active agent can be an antiparasiticagent. Non-limitative examples of antiparasitic agents are: Artemisinin,ivermectin, arteether, artemether, artenusate, eflornithine HCl,mefloquine HCl, albendazole, halofantrine, lumefantrine, quinfamide,atovaquone, bulaquine/chloroquine, and trichomonas vaccine

According to another example, the active agent can be an anticanceragent. Non-limitative examples of anticancer agents are H-101,aldesleukin, alemtuzumab, bevacizumab, celmoleukin, cetuximab,denileukin, diftitox, interferon alfa2a, interferon alfa2b, interferongamma-1a, interleukin-2, mobenakin, pegaspargase, rituximab, tasonermin,teceleukin, tositumomab, trastuzumab, aclarubicin, actinomycin D,angiotensin II, arglabin, asparaginase, bleomycin, carzinophilin,chromomycin A3, daunomycin, doxorubicin, leucovorin, masoprocol,mithramycin, mitomycin C, neocarzinostatin, paclitaxel, palictaxelnanoparticles, pentostatin, peplomycin, sarkomycin, solamargine (akaBEC), streptozocin pre-, taxotere, testosterone pre-, vinblastine,vincristine, alitretinoin, amrubicin HCl, belotecan hydrocholoride,calusterone, cladribine, cytarabine ocfosfate, dexamethasone, docetaxel,dromostanolone, elliptinium acetate, epirubicin HCl, estramustine,ethinyl estradiol pre-, toposide, exemestane, fluoxymesterone pre-,formestane, fosfestrol pre-, fulvestrant, gemtuzumab, ozogamicin,goserelin acetate, hexyl aminolevulinate, histrelin, hydroxyprogesteronepre-, idarubicin hydrochloride, irinotecan hydrochloride, leuprolide,medroxyprogesterone acetate, megesterol acetate, methylprednisolone,methyltestosterone, miltefosine, mitobronitol, nadrolonephenylpropionate, norethindrone acetate pre-, pirarubicin, prednisolonepre-, prednisone pre-, teniposide, testolactone, topotecan HCl,triamcinolone, triptorelin, valrubicin, vapreotide acetate, vindesine,vinorelbine, zinostatin stimalamer, amsacrine, arsenic trioxide,bisantrene hydrochloride, busulfan, carboplatin, carmustine (BCNU),chlorambucil, chlortrianisene pre-, cis-diamminedichloroplatinum,cyclophosphamide, dacarbazine, diethylstilbestrol pre-, flutamide,fotemustine, heptaplatin/SK-2053R, hexamethylmelamine, hydroxyurea,ifosfamide, lenalidomide, levamisole pre-, lobaplatin, lomustine (CCNU),lonidamine, mechlorethanamine, melphalan, mitotane, nedaplatin,nilutamide, nimustine hydrochloride pre-, oxaliplatin, pamidronate,pipobroman, porfimer sodium, procarbazine, ranimustine, razoxane pre-,semustine (MCCNU) pre-, sobuzoxane, sorafenib mesylate, thiotepa,triethylenemelamine pre-, zoledronic acid, anastrozole, bicalutamide,bortezomib, camostat mesylate, dasatinib, erlotinib hydrochloride,fadrozole HCl, gefitinib, imatinib mesilate, letrozole, nafoxidine pre-,sunitinib maleate, tamoxifen, toremifene, aminoglutethimide, azacytidinepre-, apecitabine, carmofur, clofarabine, cytosine arabinoside,decitabine, doxifluridine, enocitabine, floxuridine, fludarabinephosphate, fluorouracil, ftorafur, gemcitabine HCl, mercaptopurine,methotrexate, mitoxantrone HCl, nelarabine, thioguanine, uracil mustard,abarelix, bexarotene, pemetrexed, raltitrexed, tamibarotene,temozolomide, bcg live, and melanoma theraccine

According to another example, the active agent can be an antidiabeticagent. Non-limitative examples of antidiabetic agents are biphasicporcine insulin, hu neutral insulin, human insulin Zn suspension, humaninsulin zinc suspension, human neutral insulin, insulin aspart, insulinaspart/IA protamine, insulin determir, insulin glargine, insulinglulisine, insulin lispro, isophane insulin, mecasermin, oral insulin,porcine isophane insulin, porcine neutral insulin, pulmonary insulin,soluble insulin, voglibose, acarbose, extenatide, miglitol, triproamylinacetate, glimepiride, mitiglinide calcium hydrate, pioglitazone,repaglinide, epalrestat, rosiglitazone maleate, tolrestat, troglitazone,and nateglinide

According to another example, the active agent can be a natural productor natural product crude extract chosen from a vegetable, mussels (forexample, green lipped mussels), shrimps, fish, seal, microalgaee, krill,a crustacean; an hydrolysate from vegetable oil, fish oil, seal oil,microalgae oil, krill oil or crustacean oil, a vegetable oil, musselsoil (for example green lipped mussels oil), shrimps oil, fish oil, sealoil, microalgae oil, krill oil, a crustacean oil; and a proteinshydrolysate from vegetable, animal or marine source.

According to another example, the active agent can be an a naturalproduct or natural product crude extract, which can be, for example,alfalfa, aloe, angelica, arnica, aristolochic acid, artemisia,astaxanthin, ashwaganda, astragalus, avens, beta-carotene, bilberry,birch, black cohosh, black horehound, blessed thistle, biotin, boldo,burdock, calcium, calendula, california poppy, caraway, cascara sagrada,catnip, cayenne, chaste tree fruit, chondroitin sulphate, copper, crampbark, cranberry, dandelion, dang gui, devil's claw, echinacea, echinaceapurpurea, echinacea pallida, eleuthero, evening primrose oil, europeanlinden, european pennyroyal, fenugreek, feverfew, figwort, flax, folate,frankincense, garlic, gentian, ginger, ginkgo, globe artichoke,glucosamine, goldenseal, green tea, ground ivy, hawthorn, heal-all,hops, horse chestnut, horseradish, hyssop, illicium verum, juniper,licorice, linden, lungwort, lutein, melatonin, milk thistle, mugwort,niacin, pantothenic acid, peppermint, reishi mushroom, riboflavin,rosemary, saw palmetto, scullcap, selenium, schisandra, stinging nettle,St. John's wort, thuja, thyme, tomato, turmeric, valerian, willow bark,witch hazel or zeaxanthin.

According to another example, the active agent can be curcumin,artemisinin, astaxanthin, lutein, or zeaxanthin.

According to another example the active agent can be a turmeric crudeextract or purified extract from turmeric crude extract, an artemisiacrude extract or purified extract from artemisia crude extract, greenlipped mussels crude extract or purified extract from green lippedmussels crude extract, a crude extract or purified extract of amicroalgae.

According to another example the active agent can be a Haematococcuspluvialis microalgae crude extract or purified extract fromHaematococcus pluvialis microalgae crude extract.

The person skilled in the art would be able to identify, among theactive agents previously described, those that can be considered aslipophillic active agents.

According to another example, the at least one compound present in anoil can be a fatty acid or a derivative thereof (for example an C1-C6ester (C1-C6 being the amount of carbon atoms in the “alcohol” portionof the ester) of a fatty acid such as an ethyl ester) or apharmaceutically acceptable salt thereof.

According to another example, the oil can be a vegetable oil (such asflaxseed oil, pumpkinseed oil, canola oil, soybean oil, or walnut oil),fish oil (such as cod liver oil, salmon oil, tuna oil, shark oil,pelagic fishes oil, or sardine oil), seal oil, microalgae oil, krilloil, crustacean oil (for example shrimps oil), mussels oil (for examplegreen lipped mussels oil), or mixtures thereof.

The sugar can be chosen from 5-carbon sugars and 6-carbon sugars.Non-limiting examples of 5-carbon sugar include ribose, arabinose,xylose, and lyxose. Non-limiting examples of 6-carbon sugar includeglucose, galactose, mannose, allose, gulose, idose, talose, and altrose.

The sugar phosphate can be chosen from monosaccharides (such asmannose-6-phosphate, glucose-6-phosphate, galactose-6-phosphate,mannose-1-phosphate, glucose-1-phosphate and galactose-1-phosphate),disaccharides (such as6-O-phosphoryl-a-D-mannopyranosyl-(1-2)-D-mannopyranose,6-O-phosphoryl-a-D-mannopyranosyl-(1-3)-mannopyranose,6-O-phosphoryl-a-D-mannopyranosyl-(1-6)-D-mannopyranose), trisaccharides(such as6-O-phosphoryl-a-D-mannopyranosyl-(1-2)-D-mannopyranosyl-(1-2)-D-mannopyranose),and higher linear or branched oligosaccharides (such aspentamannose-6-phosphate).

The amino acid can be chosen from alanine, arginine, asparagine,aspartic acid, cysteine, glutamine, glutamic acid, glycine, isoleucine,leucine, lysine, methionine, phenylalanine, proline, serine, threonine,tryptophan, tyrosine, and valine.

The peptide can be chosen from any possible combination of the aminoacids previously described.

The compounds and compositions previously defined can be in a mixturewith a vegetable oil (such as flaxseed oil, pumpkinseed oil, canola oil,soybean oil, or walnut oil) a marine oil (such as algae oil, seal oil,krill oil, crustacean oil, or fish oil (for example cod liver oil,salmon oil, tuna oil, shark oil, pelagic fishes oil, or sardine oil,),or an hydrolysate.

The term “aryl” as used herein refers to a cyclic or polycyclic aromaticring. For example, the aryl group can be phenyl or napthyl.

The expression “aromatic heterocycle” as used herein refers to anaromatic cyclic or fused polycyclic ring system having at least oneheteroatom selected from the group consisting of N, O, S and P.Non-limitative examples include heteroaryl groups are furyl, thienyl,pyridyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, triazolyl,pyrrolyl, tetrazolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl,benzofuranyl, benzothiophenyl, carbazolyl, benzoxazolyl, pyrimidinyl,benzimidazolyl, quinoxalinyl, benzothiazolyl, naphthyridinyl,isoxazolyl, isothiazolyl, purinyl, quinazolinyl, and so on.

The expression “non-aromatic heterocycle” includes non-aromatic rings orring systems that contain at least one ring having at least one heteroatom (such as nitrogen, oxygen, sulfur or phosphorus). This termincludes, in a non-limitative manner all of the fully saturated andpartially unsaturated derivatives of the above mentioned aromaticheterocycles groups. Examples of non-aromatic heterocycle groupsinclude, in a non-limitative manner, pyrrolidinyl, tetrahydrofuranyl,morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, thiazolidinyl,isothiazolidinyl, and imidazolidinyl.

The term “crude extract” refers to an unpurified extract. Such anextract can be obtained, for example, by means of a supercritical fluidextract (for example carbon dioxide or an alkane), by using a solvent asethanol, methanol, isopropanol, acetone or water, or byhydrodistillation.

The expression “lipophilic active agent” as used herein refers to anactive agent which has an affinity for, or capability of dissolving in,lipids; i.e., non-water soluble oils, fats, sterols, triglycerides andthe like.

The term “lipid” as used herein refers to as any fat-soluble(lipophilic), molecules, such as fats, fat-like substances, oils (suchas animal oil, marine oil or vegetable oil), waxes, sterols (such ascholesterol, ergosterol, sitosterol, stigmasterol, fat-soluble vitamins(such as vitamins A, D, E and K), fatty acids, esters thereof, andvarious derivatives thereof such as monoglycerides, diglycerides,triglycerides, phospholipids, glycolipids, and cerebrosides.

The term “fatty acid(s)” as used herein refers to long chain aliphaticacids (alkanoic acids) of varying chain lengths, from about C12 to C22(although both longer and shorter chain-length acids are known). Forexample, the predominant chain lengths are about C16 to about C22. Thestructure of a fatty acid is represented by a simple notation system of“X:Y”, where X is the total number of carbon (C) atoms and Y is thenumber of double bonds.

Generally, fatty acids are classified as saturated or unsaturated. Theterm “saturated fatty acids” refers to those fatty acids that have no“double bonds” between their carbon backbone. In contrast, “unsaturatedfatty acids” are cis-isomers that have “double bonds” along their carbonbackbones. “Monounsaturated fatty acids” have only one “double bond”along the carbon backbone (e.g., usually between the 9th and 10th carbonatom as for palmitoleic acid (16:1) and oleic acid (18:1)), while“polyunsaturated fatty acids” (or “PUFAs”) have at least two doublebonds along the carbon backbone (e.g., between the 9th and 10th, and12th and 13th carbon atoms for linoleic acid (18:2); and between the 9thand 10th, 12th and 13th, and 15th and 16th for [alpha]-linolenic acid(18:3)).

“PUFAs” can be classified into two major families (depending on theposition (n) of the first double bond nearest the methyl end of thefatty acid carbon chain). Thus, the “[omega]-6 fatty acids” [omega]-6 orn-6) have the first unsaturated double bond six carbon atoms from theomega (methyl) end of the molecule and additionally have a total of twoor more double bonds, with each subsequent unsaturation occurring 3additional carbon atoms toward the carboxyl end of the molecule. Incontrast, the “[omega]-3 fatty acids” ([omega]-3 or n-3) have the firstunsaturated double bond three carbon atoms away from the omega end ofthe molecule and additionally have a total of three or more doublebonds, with each subsequent unsaturation occurring 3 additional carbonatoms toward the carboxyl end of the molecule.

The expression “inflammatory disease(s)” as used herein refers to all ofthe acute or chronic inflammatory diseases associated with the excessiverelease of cytokines, and complication thereof. The expression “chronicinflammatory disease(s)” refers to all diseases that induce tissueinjury or induce continuous inflammation due to hyperactivity and theexcessive release of cytokines, and complication thereof. In particular,the inflammatory diseases to which the compounds and compositions of thepresent disclosure can be applied are not limited to, but includeinflammatory bowel disease such as Crohn's disease and ulcerativecolitis, peritonitis, osteomyelitis, cellulitis, meningitis, cerebritis,pancreatitis, trauma-inducing shock, bronchial asthma, allergicrhinitis, cystic fibrosis, cerebral apoplexy, acute bronchitis, chronicbronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis,gout, spinal arthropathy, ankylosing spondylitis, Reiter's syndrome,psoriatic arthropathy, enteropathic spondylitis, juvenile arthropathy,juvenile ankylosing spondylitis, reactive arthropathy, infectiousarthritis, post-infectious arthritis, gonococcal arthritis, tuberculousarthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lymedisease, arthritis associated with ‘vasculitis syndrome’, polyarteritisnodosa, hypersensitivity vasculitis, Wegener's granulomatosis,polymyalgia rheumatica, giant cell arteritis, calcium crystal depositionarthropathy, pseudogout, non-joint rheumatism, bursitis, tenosynovitis,epicondylitis (tennis elbow), neuropathic joint disease(charcot joint),hemarthrosic, Henoch-Schonlein purpura, hypertrophic osteoarthropathy,multicentric reticulohistiocytoma, scoliosis, hemochromoatosis,meniscocytosis, other hemoglobinopathy, hyperlipoproteinemia,hypogammaglobulinaemia, familial mediterranean fever, Gerhardt Disease,systemic lupus erythematosus, relapsing fever, psoriasis, multiplesclerosis, sepsis (septicemia), septic shock, acute respiratory distresssyndrome, multiple organ dysfunction syndrome, chronic obstructivepulmonary disease, rheumatic arthritis, acute lung injury,bronchopulmonary dysplasia and so on.

The expression “effective amount” of a compound of the presentdisclosure or of a composition of the present disclosure is a quantitysufficient to, when administered to the subject, including a mammal, forexample a human, effect beneficial or desired results, includingclinical results, and, as such, an “effective amount” or synonym theretodepends upon the context in which it is being applied. For example, inthe context of treating cancer, for example, it is an amount of thecompound sufficient to achieve such treatment of the cancer as comparedto the response obtained without administration of the compound. Theamount of a given compound of the present disclosure that willcorrespond to an effective amount will vary depending upon variousfactors, such as the given drug or compound, the pharmaceuticalformulation, the route of administration, the type of disease ordisorder, the identity of the subject or host being treated, and thelike, but can nevertheless be routinely determined by one skilled in theart. Also, as used herein, an “effective amount” of a compound of thepresent disclosure is an amount which inhibits, suppresses or reduces acancer (e.g., as determined by clinical symptoms or the amount ofcancerous cells) in a subject as compared to a control. The samedefinition of “effective amount” also applies when the compounds of thepresent disclosure are used for inhibiting tumor growth, inhibitingtumor cell proliferation, reducing tumor growth, or enhancingbioavailability of an active agent, or treating an inflammatory disease.

The term “subject” as used herein includes all members of the animalkingdom including human. According to one embodiment, the subject is ahuman.

The expression “pharmaceutically acceptable” means compatible with thetreatment of subjects such as animals or humans.

The expression “pharmaceutically acceptable salt” means an acid additionsalt or basic addition salt which is suitable for or compatible with thetreatment of subjects such as animals or humans.

The expression “pharmaceutically acceptable acid addition salt” as usedherein means any non-toxic organic or inorganic salt of any compound ofthe present disclosure, or any of its intermediates. Illustrativeinorganic acids which form suitable salts include hydrochloric,hydrobromic, sulfuric and phosphoric acids, as well as metal salts suchas sodium monohydrogen orthophosphate and potassium hydrogen sulfate.Illustrative organic acids that form suitable salts include mono-, di-,and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic,succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic,benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonicacids such as p-toluene sulfonic and methanesulfonic acids. Either themono or di-acid salts can be formed, and such salts may exist in eithera hydrated, solvated or substantially anhydrous form. In general, theacid addition salts of the compounds of the present disclosure are moresoluble in water and various hydrophilic organic solvents, and generallydemonstrate higher melting points in comparison to their free baseforms. The selection of the appropriate salt will be known to oneskilled in the art. Other non-pharmaceutically acceptable salts, e.g.oxalates, may be used, for example, in the isolation of the compounds ofthe present disclosure, for laboratory use, or for subsequent conversionto a pharmaceutically acceptable acid addition salt. In embodiments ofthe present disclosure, the pharmaceutically acceptable acid additionsalt is the hydrochloride salt.

The term “pharmaceutically acceptable basic addition salt” as usedherein means any non-toxic organic or inorganic base addition salt ofany acid compound of the disclosure, or any of its intermediates. Acidiccompounds of the disclosure that may form a basic addition salt include,for example, where R is CO₂H. Illustrative inorganic bases which formsuitable salts include lithium, sodium, potassium, calcium, magnesium orbarium hydroxide. Illustrative organic bases which form suitable saltsinclude aliphatic, alicyclic or aromatic organic amines such asmethylamine, trimethylamine and picoline or ammonia. The selection ofthe appropriate salt will be known to a person skilled in the art. Othernon-pharmaceutically acceptable basic addition salts, may be used, forexample, in the isolation of the compounds of the disclosure, forlaboratory use, or for subsequent conversion to a pharmaceuticallyacceptable acid addition salt.

The formation of a desired compound salt is achieved using standardtechniques. For example, the neutral compound is treated with an acid orbase in a suitable solvent and the formed salt is isolated byfiltration, extraction or any other suitable method.

Compounds of the present disclosure include radiolabeled forms, forexample, compounds labeled by incorporation within the structure ²H, ³H,¹⁴C, ¹⁵N, or a radioactive halogen such as ¹²⁵I. A radiolabeled compoundof the compounds of the present disclosure may be prepared usingstandard methods known in the art.

As used herein, and as well understood in the art, “treatment” or“treating” is an approach for obtaining beneficial or desired results,including clinical results. Beneficial or desired clinical results caninclude, but are not limited to, alleviation or amelioration of one ormore symptoms or conditions, diminishment of extent of disease,stabilized (i.e. not worsening) state of disease, preventing spread ofdisease, delay or slowing of disease progression, amelioration orpalliation of the disease state, and remission (whether partial ortotal), whether detectable or undetectable. “Treatment” or “treating”can also mean prolonging survival as compared to expected survival ifnot receiving treatment.

In understanding the scope of the present disclosure, the term“comprising” and its derivatives, as used herein, are intended to beopen ended terms that specify the presence of the stated features,elements, components, groups, integers, and/or steps, but do not excludethe presence of other unstated features, elements, components, groups,integers and/or steps. The foregoing also applies to words havingsimilar meanings such as the terms, “including”, “having” and theirderivatives.

The compounds and compositions described in the present disclosure canbe useful for chemoprevention of cancer, treating cancer, inhibitingtumor growth, reducing tumor growth the prevention or treatment ofcardiovascular disease, the prevention or treatment of neurodegenerativediseases, the prevention or treatment of inflammation or of aninflammatory disease, the prevention or treatment of age-related eyedisease.

The compounds and compositions described in the present disclosure canalso be useful for in a self-emulsifying drug delivery systems (SEDDS),a self-microemulsifying drug delivery systems (SMEDDS) or aselfemulsifying oil formulations (SEOF). They can also be used for oraldelivery of lipophilic drugs, natural product or natural crude extract,as an oral bioavailability enhancer of docosahexaenoic acid,ω3-docosapentaenoic acid, ω6-docosapentaenoic acid or eicosapentaenoicacid, or for preparing a micelle or liposome for a drug delivery system.

For example, the compounds previously defined can be of formulas:

Example 1 Preparation of Monoglyceride 1

Docosapentaenoic acid ethyl ester (compound 2) (10 g) and compound 3 (6g) were mixed together and heated at a temperature of 60° C. The enzyme(100 mg) was added and the reaction mixture was stirred at 60° C. undervacuum (18 mbar) or under nitrogen bubbling for 5 h. The reactionmixture was filtered and the enzyme was washed with ethanol 95% (20 ml).The acidic resin (500 mg) or organic acid was added to the ethanolsolution and heated to reflux for 18 h. The resin was removed byfiltration and the ethanol was evaporated in vacuo. The resulting crudeproduct was dissolved in a mixture of hexanes/ethyl acetate 90:10 (10ml) and silica gel (40 g) was added. The slurry was put on a frittedfunnel and eluted with hexanes/ethyl acetate 90:10 (150 ml) to removeunreacted starting material. A second elution with ethyl acetate (300ml) give, after evaporation in vacuo, the pure compound 1 (8.7 g) wastested in vitro on the cell viability assay and in an in vivo xenografttumor model.

Pure compounds 5 and 6 (see below) have also been successfully preparedby following the same procedure.

Example 2 Preparation of a Composition (Composition 1) ComprisingVarious Monoglycerides (Compounds 1, 5 and 6)

Composition 1 comprising compounds 1, 5 and 6 was prepared according tothe same procedure as previously described in Example 1. The startingmaterial was a mixture of compounds 2, 7, and 8 at respectively (10%,80%, and 10%). This starting material composition was sold by theCompany CRODA™ Chemical Europe Ltd. under the name INCROMEGA™ DHA 700 ESR. Thus, the obtained composition 1 contains 10% of compound 1, 80% ofcompound 5, and 10% of compound 6.

Example 3

Freshly purified curcumin (10 mg) was dissolved in DMSO (1 ml). Fiveconsecutive dilutions with a equal volume of DMSO was performed. PBS wasadded to the six stock solutions to a final volume of 1% DMSO for the invitro assay. Precipitation occur at high concentration of curcumin. Tosolve the problem, curcumin (5 mg) was dissolve in a composition 1 (5mg) and DMSO (1 ml) was added. Five consecutive dilutions with a equalvolume of DMSO was performed. PBS was added to the six stock solutionsto a final volume of 1% DMSO for the in vitro assay.

The cell viability assay is performed to measure the relative cellviability status of cancer cells upon exposure to test compounds incomparison to a positive control (etoposide) and a negative control(vehicule). Adherent cells growing in 96-well plates are exposed to testcompounds for 3 days (72 hours). Four cancer cell lines including lung,colon, prostate and breast types are used since these types of cancerpossess high incidence in human. Test compounds (composition 1comprising compounds 1, 5 and 6) as well as positive and negativecontrols were tested in parallel on the same culture plate. Allconditions are tested in triplicate. Apoptotic agents such as etoposideor epigallo-catechin-gallate are used as positive controls to kill cellswhereas the solvent (dimethylsulfoxide and water) is used as negativecontrols for basal determination. Inhibition of 50% of cell growthcompared to basal condition is the lower limit indicating a positivebiological response (considered as a hit). After the incubation time,total protein content is quantified following staining with the anionicdye sulforhodamine B (SRB). The detection of luminescence, emitted bySRB, is completed by a microplate reader. This method of detection isbased upon works published by Monks et al., in Journal of the NationalCancer Institute vol. 82 no. 13 (1991) p. 757, Skehan et al. in Journalof the National Cancer Institute vol. 82 no. 13 (1990) p. 1107 andRubinstein et al. in Journal of the National Cancer Institute vol. 82no. 13 (1990) p. 1113. The amount of luminescence is directlyproportional to the number of living cells in culture.

Cancer cells were grown in T-75 flask (Falcon) containing 20 ml ofappropriate culture medium, subcultured twice a week at 37° C., 5% CO₂,95% air and 100% relative humidity and maintained at low passage number(5 to 20), following manufacturer recommendations. The cell lines usedwere A-549 (human lung carcinoma), HCT-15 (human colon adenocarcinoma),BT-549 (human breast ductal carcinoma) and PC3 (human prostateadenocarcinoma). Cells were trypsinized using 0.25% trypsine (w/v)/0.53mM EDTA solution (Hyclone), counted and plated at densities between 1000and 3000 cells per well in flat bottom 96-well clear plates (BectonDickinson) in 100 μl of appropriate culture medium supplemented withfetal bovine serum (Hyclone). Culture plates were incubated at 37° C.,5% CO₂, 95% air and 100% relative humidity for 72 hours. At 20-30% ofcell confluence, 80 μl of appropriate culture medium was added to eachwell. 20 μl of either a solution of test compounds in 6 differentsconcentration, drug for positive controls (at concentration of 29 mg/ml)or solvent (vehicle or water) for negative controls were added on top ofthe 180 μl of culture medium to obtain a final volume of 200 μl.Background plate containing the same volume of medium without cells wereincluded in each experiment. Microplates containing cells and testcompounds were incubated at 37° C., 5% CO₂, 95% air and 100% relativehumidity for 72 hours. One microplate for each cell line were fixed asdescribed below. These four microplates represented basal growth at timezero. After incubation time of 72 hours, cells were fixed with 50 μl ofcold (4° C.) 50% (w/v) trichloroacetic acid (TCA) added to the top of200 μl of culture medium. These microplates contained conditions ofgrowth control and test growth. Microplates were left 60 minutes at 4°C. and subsequently wash five times with 200 μl of deionized water.Microplates were left to dry at room temperature for at least 24 hours.All microplates were fixed with 100 μl of cold 0.4% (w/v) SRB dissolvedin 1% acetic acid solution in water added to each well containing cellsand left at room temperature for 10 minutes. Unbound SRB was removedwith successive washes (five times) with 200 μl of cold 1% acetic acidsolution in water. All microplates were left to dry at room temperaturefor at least 24 hours. Bound SRB to proteins was solubilised with theaddition of 100 μl of 10 mM cold unbuffered Tris-base solution (pH10.5). Microplates were left on a plate shaker for 5 minutes. Absorbancewas read at 515 nm using a 96-well plate Multiskan Spectrum luminescencereader (Thermo Electron Corporation). Data analysis was performed usingExcel 2003 and SigmaPlot 8.0 or GraphPadPrism 3.02 software. Percentgrowth inhibition was calculated using the absorbance measurements[Growth at time zero (T₀), growth control (C) plus the test growth atthe drug concentrations tested (T_(i)) as follows:(T_(i)−T₀)/(C−T₀)×100]. The results obtained are shown in FIGS. 1 to 9.

FIG. 1 represents the in vitro cell viability assay of six differentconcentrations of composition 1 on A-549 human lung cancer cell line.The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 25 μg/ml of the testedcomposition.

FIG. 2 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin on A-549 human lung cancer cellline. The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 100 μg/ml of curcumin.

FIG. 3 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin formulated with composition 1 onA-549 human lung cancer cell line. The positive control etoposide at 294μg/ml shows 100% growth inhibition. The 50% growth inhibition is around6.25 μg/ml of the tested composition.

Purified curcumin formulated in composition 1 decreases significantlythe concentration needed to reach the 50% growth inhibition of A-549human lung cancer cell line (see FIG. 3) as compared to purifiedcurcumin (see FIG. 2) or composition 1 alone (see FIG. 1). The 50%growth inhibition concentration of purified curcumin alone is around 100μg/ml and the 50% growth inhibition concentration of same purifiedcurcumin formulated in composition 1 is around 3.13 μg/ml, a more than30 times improvement of potency.

FIG. 4 represents the in vitro cell viability assay of six differentconcentrations of composition 1 on PC-3 human prostate cancer cell line.The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 50 μg/ml of the testedcomposition.

FIG. 5 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin on PC-3 human prostate cancer cellline. The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 75 μg/ml of curumin.

FIG. 6 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin formulated with composition 1 onPC-3 human prostate cancer cell line. The positive control etoposide at294 μg/ml shows 100% growth inhibition. The 50% growth inhibition isaround 6.25 μg/ml of the tested composition.

Purified curcumin formulated in composition 1 decreases significantlythe concentration needed to reach the 50% growth inhibition of PC-3human prostate cancer cell line (see FIG. 6) as compared to purifiedcurcumin (see FIG. 5) or composition 1 alone (see FIG. 4). The 50%growth inhibition concentration of purified curcumin alone is around 75μg/ml and the 50% growth inhibition concentration of same purifiedcurcumin formulated in composition 1 is around 3.13 μg/ml, a more than20 times improvement of potency.

FIG. 7 represents the in vitro cell viability assay of six differentconcentrations of composition 1 on HCT-15 human colon cancer cell line.The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 75 μg/ml of the testedcomposition.

FIG. 8 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin on HCT-15 human colon cancer cellline. The positive control etoposide at 294 μg/ml shows 100% growthinhibition. The 50% growth inhibition is around 100 μg/ml of curcumin.

FIG. 9 represents the in vitro cell viability assay of six differentconcentrations of purified curcumin formulated with composition 1 onHCT-15 human colon cancer cell line. The positive control etoposide at294 μg/ml shows 100% growth inhibition. The 50% growth inhibition isaround 12.5 μg/ml of the tested composition.

Purified curcumin formulated in composition 1 decreases significantlythe concentration needed to reach the 50% growth inhibition of HCT-15human colon cancer cell line (see FIG. 9) as compared to purifiedcurcumin (see FIG. 8) or composition 1 alone (see FIG. 7). The 50%growth inhibition concentration of purified curcumin alone is around 100μg/ml and the 50% growth inhibition concentration of same purifiedcurcumin formulated in composition 1 is around 6.25 μg/ml, a more than15 times improvement of potency.

Example 4

In order to determine the solubility of various compounds in a fish oilas compared to their solubility in composition 1, a first sample ofturmeric oleoresin (100 mg) obtained from ethanol extraction was stirredat room temperature in a fish oil (1.0 g) for 30 minutes. Then, anothersample of turmeric oleoresin (100 mg) (also obtained from ethanolextraction) was stirred at room temperature in composition 1 (1.0 g) for30 minutes. Both resulting suspensions were centrifuged at 12 000 RPMfor 5 minutes and 10 μl of each supernatant was dissolved in DMSO andfurther dilution was made to meet the linearity range of HPLC/MS methodfor the quantification of curcuminoids (0.001 μg/ml to 0.1 μg/ml). Thecomparative results obtained concerning the solubility of somecomponents of the turmeric oleoresin extract (bis-demethoxycurcumin,demetoxycurcumin, curcumin and total curcuminoids) in the fish oil andin the composition 1 are shown in FIG. 10.

As it can be seen in FIG. 10, composition 1 permits to enhance thesolubility of various active agents (bis-demethoxycurcumin,demetoxycurcumin, curcumin and total curcuminoids) in a lipid or lipidformulation. It can be clearly seen from FIG. 10 that such active agentshave a greater solubility in a lipid formulation consisting ofcomposition 1 than in a lipid formulation consisting of a fish oil. Asit can be seen, the active agents are about 3 to 4 times more soluble incomposition 1 than in a fish oil. It can thus also be seen the that sucha composition is effective for solubilizing lipophilic active agents.

Example 5

The relative in vivo bioavailability of two different formulationscontaining docosahexaenoic acid which are a fish oil and composition 1was determined by a pharmacokinetic study. Upon arrival in the animalfacility, the male Sprague-Dawley rats were marked for identificationand weighed. The animals were acclimatized for 1 week before commencingthe study. On the day of the study, shortly before dosing, the animalswere re-weighed and placed into experimental groups based on adistribution of weight. Animals received food and water ad libitumexcept during the pharmacokinetic study (from overnight to Bleed Time480 minutes). The animals were not deprived of food overnight from BleedTime 480 to 1440 minutes since the length of the pharmacokinetic studywas 24 hours. The compound dosing solutions were administered orally asa single slow bolus (over approximately 15 seconds) according tostandard procedures for administration of solution by gavage: the animalwas firmly restrained; a bulb-tipped gastric gavage needle of 18 G waspassed through the side of the mouth and was moved forward toward theoesophagus. The dosing solutions were dosed orally at 3 g/kg byadjusting the dose volume (3 mL/kg) according to the body weight of eachanimal and the density of the compound ˜1 g/mL). Blood samples werecollected prior to compound administration and at different time pointsfollowing administration. Blood samples (200 μl) from each animal werecollected by veinipuncture, under isoflurane anaesthesia according tothe following standard operating procedure for blood collection via thejugular vein: animals were placed in a supine position on a slantedboard to allow the head to be lower than the lower extremities. Theupper extremities were extended at a 90° angle and the neck extended andturned gently towards the site of blood collection. A 22-gauge 1″ needlewas inserted underneath the clavicle while aspirating.

For plasma preparation, blood samples were placed into tubes containingEDTA, mixed gently to assure anticoagulation and put on ice. Plasmaseparation was performed following centrifugation of the blood samples.Plasma was transferred into a tube and stored at −80° C. pendingshipment for analysis. The pellet was kept in the initial tubecontaining EDTA and stored at −80° C. pending shipment. The results ofthis study are shown in FIG. 11.

FIG. 11 shows the change in plasma docosahexaenoic acid concentration ofcomposition 1 compared to a fish oil upon time over a 1440 minutesstudy.

The relative bioavailability of docosahexaenoic acid from composition 1compared to docosahexaenoic acid from fish oil is calculated with theformula:

${{relative}\mspace{14mu}{bioavailability}} = \frac{\left\lbrack {A\; U\; C} \right\rbrack_{A}*{dose}_{B}}{\left\lbrack {A\; U\; C} \right\rbrack_{B}*{dose}_{A}}$

The AUC (calculates area under the curve for concentration vs. timedata) is calculated using linear trapezoidal rule. The use of the lineartrapezoidal rule as a method for approximating the area under aconcentration-time curve is widely accepted. In this experiment, thedoses are the same. The calculated relative bioavailability ofdocosahexaenoic acid from composition 1 compared to a fish oil from time0 to time 1440 min. is 4.48 and when the calculation is made from time 0to infinity, the relative bioavailability is 50.31.

As it can be seen in FIG. 10, composition 1 permits to enhance thesolubility of various active agents (bis-demethoxycurcumin,demetoxycurcumin, curcumin and total curcuminoids) in a lipid or lipidformulation. It can be clearly seen from FIG. 10 that such active agentshave a greater solubility in a lipid formulation consisting ofcomposition 1 than in a lipid formulation consisting of a fish oil. Asit can be seen, the active agents are about 3 to 4 times more soluble incomposition 1 than in a fish oil. It can thus also be seen the that sucha composition is effective for solubilizing lipophilic active agents.

As it can be seen in FIG. 11, composition 1 permits to enhance thebioavailability of docosahexaenoic acid as compared to a fish oil. Therelative bioavailability of docosahexaenoic acid from composition 1compared to a fish oil calculated from time 0 to time 1440 min. is 4.48and when the calculation is made from time 0 to infinity, the relativebioavailability of docosahexaenoic acid from composition 1 compared to afish oil is 50.31. Such a study thus clearly shows that the compoundsand compositions of the present disclosure are useful for enhancingbioavailability of an active agent.

Example 6

The relative human bioavailability of two different compositions(composition 2 and a fish oil) containing docosahexaenoic acid (DHA) andomega-3 docosapentaenoic acid (DPAω3) has been determined.

The fish oil comprises compounds 2 and 7 in about a 1:8 ratio (11% of 2and 89% of 7):

Composition 2 comprises compounds 1 and 5 and fish oil (comprisingcompounds 2 and 7 in about a 1:2 ratio. In other words, composition 2comprises (about) compounds 1 (3.6%), 2 (7.4%), 5 (29.4%), and 7(59.6%):

Composition 2 was prepapred according to the same procedure aspreviously described in Example 2.

The relative human bioavailability of these two different compositions(composition 2 and a fish oil) was determined by a pilot cross-overstudy on one healthy volunteer (male). The volunteer fasted for 12 hoursprior to the study. The participant consumed fish oil (capsules)equivalent to 3.0 g of DHA and 375 mg of DPAω3 as part of a breakfast.Controlled amount of boiled pasta was eaten after the 4 h time point. Aninitial blood sample (400 μl) was collected using a lancet at afingertip into heparin tubes followed by samples at 1, 2, 3, 4, 5, 6, 7and 8 hour after ingestion. Plasma was separated and immediatelyanalysed for fatty acid composition. Fourteen days later (washoutperiod), the procedure was repeated with composition 2 (capsules)equivalent to 3.0 g of DHA and 375 mg of DPAω3.

The results of this study are shown in FIGS. 12 and 13.

FIG. 12 shows the change in plasma docosahexaenoic acid (DHA)concentration of composition 2 compared to fish oil upon time over an 8hours study.

FIG. 13 shows the change in plasma omega-3 docosapentaenoic acid (DPAω3)concentration of composition 2 compared to fish oil upon time over an 8hours study.

In FIG. 12 the proportion of DHA in plasma (% DHA) increased slowly onlyafter 3 hours and reach a maximum of less than 2% after 8 hours whenfish oil was taken alone. With composition 2, the DHA increasedmoderately right after the ingestion and after 4 hours the DHA increasedrapidly to reach a plateau of more than 4.5% at 6 hours. After 8 hoursthe DHA variation is 4.5%.

In FIG. 13 the proportion of DPAω3 in plasma (% DPAω3) did not increaseafter 8 hours when fish oil was taken alone, this mean that DPAω3 wasnot absorbed in fish oil. With composition 2, the DPAω3 increasedmoderately right after the ingestion to reach a plateau of 0.5% at 3hours. After 8 hours the DPAω3 variation was more than 0.6%

The relative bioavailability of fatty acids from composition 2 comparedto fish oil is calculated with the formula:

${{relative}\mspace{14mu}{bioavailability}} = \frac{\left\lbrack {A\; U\; C} \right\rbrack_{A}*{dose}_{B}}{\left\lbrack {A\; U\; C} \right\rbrack_{B}*{dose}_{A}}$

The AUC (calculates area under the curve for concentration vs. timedata) is calculated using linear trapezoidal rule. The use of the lineartrapezoidal rule as a method for approximating the area under aconcentration-time curve is widely accepted. In this experiment, thedoses are the same. The calculated relative bioavailability ofdocosahexaenoic acid from composition 2 compared to fish oil from time 0to infinity is 3.72. Thus, when DHA is in the presence of compounds 1and/or 5, DHA is 3.72 times more bioavailable. For the relativebioavailability of DPAω3, no significant absorption was found with fishoil, compared to an increase of more than 0.6% after 8 hours withcomposition 2. The relative bioavailability of compound 1 and compound 5is calculated with the same formula:

${{relative}\mspace{14mu}{bioavailability}} = \frac{\left\lbrack {A\; U\; C} \right\rbrack_{A}*{dose}_{B}}{\left\lbrack {A\; U\; C} \right\rbrack_{B}*{dose}_{A}}$

The calculated relative bioavailability of compound 1 compared tocompound 5 from time 0 to infinity is 2.20. Thus, compound 1 is 2.2times more bioavailable than compounds 5.

The compounds and compositions of the present disclosure can be used forenhancing bioavailability of an active agent. For example, the activeagent can be a fatty acid or a derivative thereof (for example an esterof a fatty acid). For example, the compounds of the present disclosurecan be used for enhancing bioavailability of at least one compoundpresent in a fish oil. For example, the compounds of the presentdisclosure can be used for enhancing bioavailability of the ethyl esterof EPA and/or DHA.

Example 7

Composition 3 (comprising compounds 1 and 5) at final concentration of10 μg/ml, curcumin (5 μg/ml) and a 1:1 mixture of composition 3 (10μg/ml) and curcumin (5 μg/ml) in DMSO (1%) was used for the in vitroassay. Composition 3 prepared according to the same procedure aspreviously described in Example 2)

The in vitro assay allows evaluation of the potential anti-inflammatoryeffects of compounds on the induced-release of pro-inflammatory mediatorby monocyte cells. Typical human monocyte THP-1 cells, involved ininflammatory processes, are used in this assay. Measurement ofpro-inflammatory mediator TNF-α is performed by ELISA (manufactured byR&D Systems) with artificial induction of pro-inflammatory agents by LPS(E. Coli O55:B5) during 4 hours. Known anti-inflammatory agentdexametazone was used as positive control.

The results of this study are shown in FIG. 14.

In FIG. 14, no TNF-α was measured when no LPS is added to the monocyteTHP-1 cells incubated with compounds or vehicle. With 100 ng/ml of LPS,400 pg/ml of TNF-α was measured with the vehicle. With positive controldexametazone, only 125 pg/ml of TNF-α was measured, showing theanti-inflammatory effect of dexametazone. When composition 3 (10 μg/ml)was added, 275 pg/ml of TNF-α was measured and 100 pg/ml of TNF-α wasmeasured when curcumin (5 μg/ml) is added. When a mixture of composition3 and curcumin was added, less than 50 pg/ml of TNF-α was measured,showing a strong anti-inflammatory synergic effect.

While the compounds, compositions, methods and uses thereof have beendescribed in connection with specific embodiments thereof, it will beunderstood that they can be further modified and this application isintended to cover any variations, uses, or adaptations of the compounds,compositions, methods and uses thereof following, in general, theprinciples described in the present disclosure and including suchdepartures from the present disclosure as come within known or customarypractice within the art to which the present disclosure pertains and asmay be applied to the features hereinbefore set forth, and as follows inthe scope of the appended claims.

What is claimed is:
 1. A method for treating lung cancer, prostatecancer, breast cancer or colon cancer comprising administering to asubject in need thereof an effective amount of curcumin and an effectiveamount of at least one compound chosen from compounds of formulae (I),(II), (III), and (IV):

wherein in formula (I): X₁ is O, NH, or S; X₂ is O, NH, or S; X₃ is O,NH, or S; R₁ and R₂ each independently represents —H, —C(O)NH₂,—S(O)NH₂, or —S(O)₂NH₂; in formula (II): X₁ is O, NH, or S; X₂ is O, NH,or S; X₃ is O, NH, or S; R₁ and R₂ each independently represents —H,—C(O)NH₂, —S(O)NH₂, or —S(O)₂NH₂; in formula (III) X₁ is O, NH, or S; X₂is O, NH, or S; X₃ is O, NH, or S; R₁ and R₂ each independentlyrepresents, —H, —C(O)NH₂, —S(O)NH₂, or —S(O)₂NH₂; in formula (IV) X₁ isO, NH, or S; X₂ is O, NH, or S; X₃ is O, NH, or S; R₁ and R₂ eachindependently represents, —H, —C(O)NH₂, —S(O)NH₂, or —S(O)₂NH₂, or apharmaceutically acceptable salt thereof.
 2. The method of claim 1,wherein a composition comprising an effective amount of curcumin and aneffective amount of at least one compound chosen from compounds offormulae (I), (II), (III), and (IV) is administered to said subject. 3.The method of claim 1, wherein X₁ is O, X₂ is O, and X₃ is O.
 4. Themethod of claim 3, wherein R₁ is —H and R₂ is —H.
 5. The method of claim1, wherein X₁ is NH, X₂ is O, and X₃ is O.
 6. The method of claim 5,wherein R₁ is —H and R₂ is —H.
 7. The method of claim 1, wherein X₁ isO, X₂ is NH, and X₃ is O.
 8. The method of claim 7, wherein R₁ is —H andR₂ is —H.
 9. The method of claim 1, wherein X₁ is O, X₂ is O, and X₃ isNH.
 10. The method of claim 9, wherein R₁ is —H and R₂ is —H.
 11. Themethod of claim 4, wherein said at least one compound is a compound offormula (IV).
 12. The method of claim 4, wherein said at least onecompound is a compound of formula (IV).
 13. The method of claim 1,wherein said effective amount of curcumin and said effective amount ofsaid at least one compound are administered separately.
 14. The methodof claim 4, wherein said effective amount of curcumin and said effectiveamount of said at least one compound are administered separately. 15.The method of claim 1, wherein said cancer is prostate cancer.
 16. Themethod of claim 1, wherein said cancer is breast cancer.
 17. The methodof claim 1, wherein said cancer is lung cancer.
 18. The method of claim1, wherein said cancer is colon cancer.
 19. The method of claim 4,wherein said cancer is prostate cancer.
 20. The method of claim 4,wherein said cancer is breast cancer.
 21. The method of claim 4, whereinsaid cancer is lung cancer.
 22. The method of claim 4, wherein saidcancer is colon cancer.
 23. The method of claim 1, wherein X₁ is O, orNH; X₂ is O, or NH; and X₃ is O, or NH.
 24. The method of claim 23,wherein R₁ is —H and R₂ is —H.